American Society of Hematology

Figure 2

$Figure 2. Malformations and blood perfusion (the “nonseparation” phenotype) of cutaneous lymphatic vessels in podoplanin−/− mice. (A) In a wild-type P3, mouse the en face view of the dermal side of the skin of the flanks and abdomen shows blood-filled veins of different calibers. The lymphatic system is visualized by VEGFR-3–driven, lymphatic endothelium–specific expression of β-galactosidase (inset). (B) By contrast, in P3 podoplanin−/− mice, a fractal pattern of blood-filled, tortuous, blind-ending vessels are found, besides occasional areas of bleeding (white asterisk). These vessels are of lymphatic origin because they express VEGFR-3–driven β-galatosidase (inset); Olympus SZ zoom stereo microscope, Sony DSC W200 camera with adapter VAE-WD. (C) Sections through the flank skin of P3 podoplanin+/+ mice show Lyve-1–expressing lymphatics (brown) in the upper dermis. These vessels also express Prox-1 (red) on top of Lyve-1 (green) by double immunofluorescence (inset). (D) Skin of age-matched podoplanin−/− mice contains extended, randomly distributed lymphatic vessels. Their endothelial cells express Prox-1 (red) and Lyve-1 (green), and their dilated lumen contains erythrocytes (yellow) that also have leaked into the perivascular space (white arrowhead; inset). (E) In a P3 podoplanin−/− mouse, intravenously injected Chicago sky blue fills dermal veins and arteries (“purple vessel” on the left side of the image; Olympus SZ40); however, the blood-filled lymphatics (green arrowheads) are not entered by the tracer, indicating that this compartment is already sequestered from the circulation in this P3 mouse. Identification of perfused lymphatic vessels (F) by intravenous injection of FITC-lectin (green). Lyve-1 is blue and Ter119 is red. FITC-lectin is not detectable in the small erythrocyte-filled lymphatic capillaries (arrow in panel G). (H) Control injection of FITC-lectin into wild-type mice. FITC-lectin is confined to blood vessels and not detectable in large (arrow) or small lymphatic vessels (the latter are not shown in this picture). Scale bars in panels A, B, and E equal 300 μm; in panels C and D, 100 μm; in panel D inset, 15 μm; and in panels F through H, 50 μm.$

Malformations and blood perfusion (the “nonseparation” phenotype) of cutaneous lymphatic vessels in podoplanin^−/− mice. (A) In a wild-type P3, mouse the en face view of the dermal side of the skin of the flanks and abdomen shows blood-filled veins of different calibers. The lymphatic system is visualized by VEGFR-3–driven, lymphatic endothelium–specific expression of β-galactosidase (inset). (B) By contrast, in P3 podoplanin^−/− mice, a fractal pattern of blood-filled, tortuous, blind-ending vessels are found, besides occasional areas of bleeding (white asterisk). These vessels are of lymphatic origin because they express VEGFR-3–driven β-galatosidase (inset); Olympus SZ zoom stereo microscope, Sony DSC W200 camera with adapter VAE-WD. (C) Sections through the flank skin of P3 podoplanin^+/+ mice show Lyve-1–expressing lymphatics (brown) in the upper dermis. These vessels also express Prox-1 (red) on top of Lyve-1 (green) by double immunofluorescence (inset). (D) Skin of age-matched podoplanin^−/− mice contains extended, randomly distributed lymphatic vessels. Their endothelial cells express Prox-1 (red) and Lyve-1 (green), and their dilated lumen contains erythrocytes (yellow) that also have leaked into the perivascular space (white arrowhead; inset). (E) In a P3 podoplanin^−/− mouse, intravenously injected Chicago sky blue fills dermal veins and arteries (“purple vessel” on the left side of the image; Olympus SZ40); however, the blood-filled lymphatics (green arrowheads) are not entered by the tracer, indicating that this compartment is already sequestered from the circulation in this P3 mouse. Identification of perfused lymphatic vessels (F) by intravenous injection of FITC-lectin (green). Lyve-1 is blue and Ter119 is red. FITC-lectin is not detectable in the small erythrocyte-filled lymphatic capillaries (arrow in panel G). (H) Control injection of FITC-lectin into wild-type mice. FITC-lectin is confined to blood vessels and not detectable in large (arrow) or small lymphatic vessels (the latter are not shown in this picture). Scale bars in panels A, B, and E equal 300 μm; in panels C and D, 100 μm; in panel D inset, 15 μm; and in panels F through H, 50 μm.

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